Presented here is the complete genome sequence of Thiomicrospira crunogena XCL-2, representative of ubiquitous chemolithoautotrophic sulfur-oxidizing bacteria isolated from deep-sea hydrothermal vents. This gammaproteobacterium has a single chromosome (2,427,734 base pairs), and its genome illustrates many of the adaptations that have enabled it to thrive at vents globally. It has 14 methyl-accepting chemotaxis protein genes, including four that may assist in positioning it in the redoxcline. A relative abundance of coding sequences (CDSs) encoding regulatory proteins likely control the expression of genes encoding carboxysomes, multiple dissolved inorganic nitrogen and phosphate transporters, as well as a phosphonate operon, which provide this species with a variety of options for acquiring these substrates from the environment. Thiom. crunogena XCL-2 is unusual among obligate sulfur-oxidizing bacteria in relying on the Sox system for the oxidation of reduced sulfur compounds. The genome has characteristics consistent with an obligately chemolithoautotrophic lifestyle, including few transporters predicted to have organic allocrits, and Calvin-Benson-Bassham cycle CDSs scattered throughout the genome.

Bacillus anthracis spores have been used as a biological weapon in the United States. We wanted to develop a safe, rapid method of sample preparation that provided safe DNA for the detection of spores in environmental and clinical specimens. Our method reproducibly detects B. anthracis in samples containing <10 spores.Bacillus anthracis spores, a recent threat, can remain dormant for years while retaining full virulence (5,10,13,20,21,22; S. Endicott, E. Hagerman, and M. Furmanski, Letter, JAMA 284:561-562, 2000). Powders and environmental samples, the most common nonclinical specimens, and nasopharyngeal swabs are submitted to designated laboratories in the national Laboratory Response Network (LRN), which has facilities to work safely on the specimens (6,19,23). High numbers of specimen can overwhelm a laboratory's capability and capacity to perform the tests in a timely fashion. From October to December 2001, the Florida Department of Health's three LRN laboratories each received hundreds of samples per day, which revealed the need for a method to render the samples harmless so the DNA could be safely extracted under biological safety level 2 conditions, to alleviate the bottleneck, and to decrease the turnaround time to the final result. Specimens may contain Ͻ10 spores, and the isolation and identification of B. anthracis may take days (5, 15). The purpose of this work was to develop a method of sample preparation that would provide safe DNA for the detection of Յ10 B. anthracis spores.Previous studies used sonication probes in open tubes and cartridges to hold samples, a method that necessitated the use of a biological safety level 3 environment and sterilization between each sample preparation (2, 3, 7). Dang et al. demonstrated that DNA from autoclaved spores was usable for PCR assays (9). No one explored the sensitivity of these methods or if combinations of methods could be used.(A preliminary report of this work has been presented previously [V. A. Luna, M.

In this study, food samples were intentionally contaminated with Escherichia coli O157:H7, and then DNA was isolated by using four commercial kits. The isolated DNA samples were compared by using real-time PCR detection of the Shiga toxin genes. The four kits tested worked similarly.

Molecular methods have become vital epidemiological tools in the detection and characterization of bacteria associated with a foodborne outbreak. We used both culture and real-time PCR to detect a Vibrio parahaemolyticus isolate associated with a foodborne outbreak. The outbreak occurred in July 2002 in Polk County, Florida, and originated at a Chinese buffet, with one person being hospitalized. The hospital isolated V. parahaemolyticus from the patient but destroyed the sample after diagnosis. From an onsite visit of the restaurant, food samples that possibly contributed to the outbreak were collected and sent to the Florida Department of Health, Tampa Branch Laboratory. Crab legs, crabsticks, and mussel samples were homogenized and incubated according to the Food and Drug Administration Bacteriological Analytical Manual culture protocol. Three sets of primers and a TaqMan probe were designed to target the tdh, trh, and tlh genes and used for real-time PCR. This study was successful in isolating V. parahaemolyticus from a mussel sample and detecting two of its genes (tdh and tlh) in food and pure culture by real-time PCR.